Protozoans of the Leishmania genus are the etiological agents of a wide spectrum of diseases commonly known as leishmaniases. Lipid organization of the plasma membrane of the parasite may mimic the lipid organization of mammalian apoptotic cells and play a role in phagocytosis and parasite survival in the mammal host. Here, we analyzed the phospholipid dynamics in the plasma membrane of both the L. (Leishmania) and the L. (Viannia) subgenera. We found that the activity and substrate specificity of the inward translocation machinery varied between Leishmania species. The differences in activity of inward phospholipid transport correlated with the different sensitivities of the various species towards the alkyl-phospholipid analogue miltefosine. Furthermore, all species exhibited a phospholipid scramblase activity in their plasma membranes upon stimulation with calcium ionophores. However, binding of annexin V to the parasite surface was only detected for a subpopulation of parasites during the stationary growth phase and only marginally enhanced by scramblase activation.
References
[1]
Cunningham AC (2002) Parasitic adaptive mechanisms in infection by leishmania. Exp Mol Pathol 72: 132–141.
[2]
Lainson R, Shaw JJ (1987) Evolution, classification and geographical distribution. In: Peters W, Killick-Kendrick R, editors. The Leishmaniases in Biology and Medicine.London: Academic Press Inc. 1–120.
[3]
de Freitas Balanco JM, Moreira ME, Bonomo A, Bozza PT, Amarante-Mendes G, et al. (2001) Apoptotic mimicry by an obligate intracellular parasite downregulates macrophage microbicidal activity. Curr Biol 11: 1870–1873.
[4]
van Zandbergen G, Bollinger A, Wenzel A, Kamhawi S, Voll R, et al. (2006) Leishmania disease development depends on the presence of apoptotic promastigotes in the virulent inoculum. Proc Natl Acad Sci U S A 103: 13837–13842.
[5]
Wanderley JL, Pinto da Silva LH, Deolindo P, Soong L, Borges VM, et al. (2009) Cooperation between apoptotic and viable metacyclics enhances the pathogenesis of Leishmaniasis. PLoS One 4: e5733.
[6]
Zhang K, Pompey JM, Hsu FF, Key P, Bandhuvula P, et al. (2007) Redirection of sphingolipid metabolism toward de novo synthesis of ethanolamine in Leishmania. EMBO J 26: 1094–1104.
[7]
Zheng L, T'Kind R, Decuypere S, von Freyend SJ, Coombs GH, et al. (2010) Profiling of lipids in Leishmania donovani using hydrophilic interaction chromatography in combination with Fourier transform mass spectrometry. Rapid Commun Mass Spectrom 24: 2074–2082.
[8]
Zufferey R, Allen S, Barron T, Sullivan DR, Denny PW, et al. (2003) Ether phospholipids and glycosylinositolphospholipids are not required for amastigote virulence or for inhibition of macrophage activation by Leishmania major. J Biol Chem 278: 44708–44718.
[9]
Weingartner A, Kemmer G, Muller FD, Zampieri RA, Gonzaga dos Santos M, et al. (2012) Leishmania promastigotes lack phosphatidylserine but bind annexin V upon permeabilization or miltefosine treatment. PLoS One 7: e42070.
[10]
Yeung T, Heit B, Dubuisson JF, Fairn GD, Chiu B, et al. (2009) Contribution of phosphatidylserine to membrane surface charge and protein targeting during phagosome maturation. J Cell Biol 185: 917–928.
[11]
Holthuis JC, Levine TP (2005) Lipid traffic: floppy drives and a superhighway. Nat Rev Mol Cell Biol 6: 209–220.
[12]
Devaux PF, Lopez-Montero I, Bryde S (2006) Proteins involved in lipid translocation in eukaryotic cells. Chem Phys Lipids 141: 119–132.
[13]
Smeets EF, Comfurius P, Bevers EM, Zwaal RF (1994) Calcium-induced transbilayer scrambling of fluorescent phospholipid analogs in platelets and erythrocytes. Biochim Biophys Acta 1195: 281–286.
[14]
Williamson P, Bevers EM, Smeets EF, Comfurius P, Schlegel RA, et al. (1995) Continuous analysis of the mechanism of activated transbilayer lipid movement in platelets. Biochemistry 34: 10448–10455.
[15]
Sims PJ, Wiedmer T (2001) Unraveling the mysteries of phospholipid scrambling. Thromb Haemost 86: 266–275.
[16]
Araujo-Santos JM, Gamarro F, Castanys S, Herrmann A, Pomorski T (2003) Rapid transport of phospholipids across the plasma membrane of Leishmania infantum. Biochem Biophys Res Commun 306: 250–255.
[17]
Tripathi A, Gupta CM (2003) Transbilayer translocation of membrane phosphatidylserine and its role in macrophage invasion in Leishmania promastigotes. Mol Biochem Parasitol 128: 1–9.
[18]
Weingartner A, Drobot B, Herrmann A, Sanchez-Canete MP, Gamarro F, et al. (2010) Disruption of the lipid-transporting LdMT-LdRos3 complex in Leishmania donovani affects membrane lipid asymmetry but not host cell invasion. PLoS One 5: e12443.
[19]
Perez-Victoria FJ, Sanchez-Canete MP, Castanys S, Gamarro F (2006) Phospholipid translocation and miltefosine potency require both L. donovani miltefosine transporter and the new protein LdRos3 in Leishmania parasites. J Biol Chem 281: 23766–23775.
[20]
Weingartner A, Dos Santos MG, Drobot B, Pomorski TG (2011) Ca(2+)-activated transbilayer movement of plasma membrane phospholipids in Leishmania donovani during ionomycin or thapsigargin stimulation. Mol Biochem Parasitol
[21]
Parodi-Talice A, Araujo JM, Torres C, Perez-Victoria JM, Gamarro F, et al. (2003) The overexpression of a new ABC transporter in Leishmania is related to phospholipid trafficking and reduced infectivity. Biochim Biophys Acta 1612: 195–207.
[22]
Araujo-Santos JM, Parodi-Talice A, Castanys S, Gamarro F (2005) The overexpression of an intracellular ABCA-like transporter alters phospholipid trafficking in Leishmania. Biochem Biophys Res Commun 330: 349–355.
[23]
Castanys-Munoz E, Alder-Baerens N, Pomorski T, Gamarro F, Castanys S (2007) A novel ATP-binding cassette transporter from Leishmania is involved in transport of phosphatidylcholine analogues and resistance to alkyl-phospholipids. Mol Microbiol 64: 1141–1153.
[24]
Castanys-Munoz E, Perez-Victoria JM, Gamarro F, Castanys S (2008) Characterization of an ABCG-like transporter from the protozoan parasite Leishmania with a role in drug resistance and transbilayer lipid movement. Antimicrob Agents Chemother 52: 3573–3579.
[25]
Weingartner A, dos Santos MG, Drobot B, Pomorski TG (2011) Ca2+-activated transbilayer movement of plasma membrane phospholipids in Leishmania donovani during ionomycin or thapsigargin stimulation. Mol Biochem Parasitol 179: 59–68.
[26]
Wanderley JL, Moreira ME, Benjamin A, Bonomo AC, Barcinski MA (2006) Mimicry of apoptotic cells by exposing phosphatidylserine participates in the establishment of amastigotes of Leishmania (L) amazonensis in mammalian hosts. J Immunol 176: 1834–1839.
[27]
Perez-Victoria FJ, Gamarro F, Ouellette M, Castanys S (2003) Functional cloning of the miltefosine transporter. A novel P-type phospholipid translocase from Leishmania involved in drug resistance. J Biol Chem 278: 49965–49971.
[28]
Alder-Baerens N, Lisman Q, Luong L, Pomorski T, Holthuis JC (2006) Loss of P4 ATPases Drs2p and Dnf3p disrupts aminophospholipid transport and asymmetry in yeast post-Golgi secretory vesicles. Mol Biol Cell 17: 1632–1642.
[29]
Chen S, Wang J, Muthusamy BP, Liu K, Zare S, et al. (2006) Roles for the Drs2p-Cdc50p complex in protein transport and phosphatidylserine asymmetry of the yeast plasma membrane. Traffic 7: 1503–1517.
[30]
Farias LH, Rodrigues AP, Silveira FT, Seabra SH, Damatta RA, et al. (2012) Phosphatidylserine exposure and surface sugars in two Leishmania (Viannia) braziliensis strains involved in cutaneous and mucocutaneous leishmaniasis. J Infect Dis
[31]
Howard MK, Pharoah MM, Ashall F, Miles MA (1991) Human urine stimulates growth of Leishmania in vitro. Trans R Soc Trop Med Hyg 85: 477–479.
[32]
Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–917.
[33]
Spath GF, Beverley SM (2001) A lipophosphoglycan-independent method for isolation of infective Leishmania metacyclic promastigotes by density gradient centrifugation. Exp Parasitol 99: 97–103.
[34]
Saraiva EM, Pinto-da-Silva LH, Wanderley JL, Bonomo AC, Barcinski MA, et al. (2005) Flow cytometric assessment of Leishmania spp metacyclic differentiation: validation by morphological features and specific markers. Exp Parasitol 110: 39–47.